Compounds of the cyclopentanopolyhydrophenanthrene series and a method for producing them



Patented May 26, 1942 COMPOUNDS OF THE CYCLOPENTANOPOLY-HYDROPHENANTHEENE SERIES AND A METHOD FOR PRODUCING THEM AdolfButenandt, Danzig-Langtuhr, Free State of Danzig, assignor to ScheringCorporation, Bloomfield, N. a corporation of New Jersey No Drawing.Application November 26, 1934, Serial No. 754,854. In Germany November25,

22 Claims. (01. 260-4914) (Granted under the provisions of sec. 14, actof March 2, 1927; 357 0. G. 5)

This invention relates to the isolation of the male sex hormone from itssources, and more particularly to the conversion of by-products oraccompanying substances obtained in the course of the known processesofproducing the male .sex hormone, and containing the same carbon skeletonas the hormone, into the latter or its derivatives and therapeuticallyvaluable unsaturated hydroaromatic ketones of a higher order, i. e.,ketoclyopentanodimethylpolyhydrophenanthroles.

It is already known to separate the male sex hormone of the formulaCmHaoOz in crystalline form from extracts of the urine of maleindividuals. But the yields obtained thereby are very small. Besides,larger amounts of other compounds are found which, hitherto, have notbeen isolated or made use of. These accompanying compounds have the samecarbon skeleton as androsterone but are of unsaturated nature and maycontain substituents in the same or in sterically diflerent manner fromandrosterone, among such compounds being transdehydro-androsterone.

We have found that the male sex hormone or its derivatives can besynthesized from these hitherto useless compounds. For instance, whenChemischen Analyse, by Hoppe; Seyler-Thierfelder, 9th ed. p. 327, Berlin1924), this compound gives a roseate coloration.

starting with a petroleum ether extract as it remains according to theexample of the copending application Serial No. 638,390, filedOctober18, 1932, which has matured into Patent No. 2,015,099, after separatingthe male sex hormone by treating with 60% alcohol, and treating saidextract with higher concentrated alcohol, for instance with 80% alcohol,or with other water-miscible solvents of suitable concentration, forinstance with methanol and the like (the optimum amount of water can ineach case be determined by simple experiment), a further portion havingalso ketonic functions, passes into said solvent. When adding reagentsfor the keto group to this solution, such as semicarbazide,hydroxylamine, phenyl hydrazine and the like, a semicarbazone, .oxime,phenyl hydrazone or the like crystallizes which, on splitting withdilute sulfuric acid in alcoholic solution, yields on further working upa white crystalline com; pound of the melting point 156.5-157.5 C. Itcan be purified by recrystallization from methanol. Its analysescorrespond to the formula Ciel-127001. Besides one ketogroup, itcontains one atom of chlorine; furthermore, the presence of a doublebond in its molecule can be proved; for, on testing according toLiebermann-Burk- Also other starting materials than the urine of maleindividuals can be used for isolating this and similar chloro ketoneswhen said starting materials were previously subjected to the action ofhydrochloric acid. In the place of hydrochloric acid, also otherhydrohalic acids or agents capable of yielding hydrogen halide, can beused. One may also proceed in such a manner that the crude extractsobtained according to Funk (Proceedings of the Society for ExperimentalBiology and Medicine, 26, page 569 (1929)) or crude products obtainedsynthetically, for instance, by the oxidation of sterines, are directlyworked up to such unsaturatedchloroketones. It is also possible tosubject the entire residue directly without preliminary treatment withsolvents to the treatment with reagents for keto groups and to work upthe reaction products obtained in the described manner.

The double bond in such unsaturated ketones can be readily hydrogenatedby activated hydrogen, for instance by the action of hydrogen in thepresence of catalysts, such as palladium and the like, the keto groupremaining practically unattacked by this treatment. The

hydrogenation can-also be carried out at an elevated temperature. Thereaction may also take place under pressure. Besides palladium, alsoother customary hydrogenation catalysts or their mixtures can be used ascatalysts. Furthermore in the place of catalytically activated hydrogen,hydrogen which has been activated in another manner, for instancehydrogen in statu nascendi, can be employed.

The hydrogenation of the mentioned chloroketone of the melting point1565-1575 C. is

carried out, for instance, in such a manner that the chloroketone isdissolved in alcohol and after the addition of colloidal palladium isshaken with hydrogen. The reaction proceeds more rapidly at an elevatedtemperature. When the absorption of hydrogen has come to an end, the

solution is filtered oflf from the catalyst and is concentrated. Therebya saturated chloroke tone of the formula C19H29OC1 with a melting pointof 173 C. crystallizes which does not give any of the color reactions ofthe unsaturated ketone.

The conversion of said saturated chloroketone into the male sex hormonerequires the substitution of the chlorine atom by a hydroxy ard.(Handbuch der Physlologisch-Pathologischgroup. This may, for instance,be achieved in such a manner that first the ester derivative is producedby reacting the chloroketone, according to the principle of the doubleconversion,

with salts of carboxylic or sulfonic acids which ester derivatives arethen saponifled, if requlred. However, one may also employ other methodsfor the conversion of thechlorine atom -quently, areextracted ten timeswith about 100. cc. of 85% alcohol, (denatured with methanol). Thecombined alcoholic portions, after evaporation anddrylng. leave about.10 g. of a residue which still contains some alcohol which is heatedwith a solution of 5 g. of semicarbazide acetate in into the OHS-group,as for instance, by directly exchanging them or by introducingintermediate substituents such as the amino group.

isolated in white needles by fractional crystallization from methanol.By saponifying the acetate, the known male sex hormone in purecrystalline form is produced.

Instead of potassium acetate, also other acetates, for instance silveracetate, or, in general, the salts of carboiwlio and sulfonic acids canbe used. The isolation of the hormone esters ob tained can also becarried out by fractionated sublimation in a high vacuum.

Under the conditions of the production of the In order to carry out thisconversion reactionabove mentioned hormone esters from the halogenketone there are formed, due to the splitting of! of hydrogen halide,unsaturated ketones as by-products. These are separated from the hormoneesters by fractional crystallization or sublimation in a high vacuum. Itwas possible to produce from this mixture of unsaturated hydroaromaticketones a product of the melting point 103 C. in rectangular lamellas(from methanol). The double bond in this product can be demonstrated bytitration with per benzoic acid as well as by the addition of bromine.The dibromide of this compound can be obtained in crystalline form fromalcohol.

0! course, one may also proceed in such a manner that the unsaturatedhalogen ketone is first transformed into the ester and then hydrogenatedand, if required, saponifled, or that the esterified unsaturated ketoneis first saponitied and then hydrogenated.

The process described enables one to produce the saturated as well asthe unsaturated male sex hormone or its derivatives in a convenientmanner from hitherto useless accompanying substances by-products fromthe isolation of the same.

The following examples serve to illustrate the invention without,however, limiting the same to them:

' Example 1 30 grams of a "petroleum ether portion" obtained on workingup extracts produced from about 4,000 liters of the urine of maleindividuals, to androsterone, as described, for example, in the abovementioned patent, said portion being practically without anyphysiological activity, are digested with 4 cc. of benzene on the waterbath,

70 cc. of absolute alcohol for about 12 hours on a water bath sogently'that the reaction mass is.

insplssated during this time to fa syrupy consistency. After-cooling, itis triturated with a little cooled alcohol, is filtered off by suctionand the crystal paste collected on the filter, and 'is alternatelywashed with cooled alcohol, ether and hot water. The crudesemicarbazone-obtained thereby represents a yellowish crystallinepowder.

The decomposition point lies at about 240 C.

The crude semicarbazone can be recrystallized by extraction withpropanol in a Soxhlet, apparatus and is obtained in white laminas ofthemelting point 275 C. For the purpose of further working up the. crudesemicarbazone, 1g. of the same is dissolved, while stirring, in 35 cc.of dilute alcoholic sulfuric acid (10 cc, of concentrated sulfuric acid,15 cc. of water, 75 cc. of alcohol) or in 35 cc. of an alcoholic-aqueoushydrochloric acid (50 cc. of concentrated hydrochloric acid, 50cc. ofalcohol) and is heated for 15 to 20 minutes on the boiling water bath.After diluting with water, it is extracted with ether, the etherealsolution is washed with bicarbonate and water,

is dried and evaporated to dryness. The crude cleavage product (about0.63 g.) is triturated with very little freshly distilled ethanol and iscaused to crystallize by cooling. The chloroketone which precipitates isrecrystallized from freshly distilled methanol until it shows theconstant melting point of 156.5-157.5 C. (not corrected); it is readilysoluble in ether, acetone, chloroform, dioxane acetic acid ester,soluble in petroleum ether, d iflicultly soluble in cold, readily in hotalcohol. The chloroketone forms elongated mixed with 300 cc. ofpetroleum ether and fil prisms which .sublimate from 110 C. on at a tionwith tetranitro methane, decolorizes a bromine-glacial acetic acidsolution, exhibits a positive Beilstein test and in acetic acidanhydride with concentrated sulfuric acid a yellow coloration whichrapidly changes to intensively pink The oxime has a melting point of168-169 C.

The chloroketone can be readily hydrogenated in alcoholic or in glacialacetic acid solution when using a palladium catalyst (palladium black,palladium-barium sulfate or palladiumcalcium carbonate catalyst). Afterone double bond is saturated. also the carbonylic group is attacked andis surprisingly readily reduced to the CHOH- or the CH2-- group; forthis reason it proves to be expedient to precipitate the saturatedohloroketone in the form of the semicarbazone (melting point 285 C.)from the mother liquors of which those portions of the hydrogenationproducts which are richer in hydrogen, can be obtained by sublimation ina high vacuum.

By decomposing the semicarbazone by means of dilute acids, the dihydrochloroketone CroHsoOCl is obtained which crystallizes from dilutealcohol in prismatic needles of the melting point 173 0. (notcorrected), and which yields with tetranitro methane no color reactionat all and with acetic acid anhydride and concentrated sulfuric acidonly a slightly yellow coloration. Yield up to The reaction of thedihydro chloroketone of melting point 173 C. with potassium acetate wascarried out by heating. the halogen compound with a mixture of potassiumacetate and glacial acetic acid for hours in a bomb tube to about 160 C.The reagent was prepared in the following manner: Purest potassiumacetate was dissolved in hot glacial acetic acid; the solution wasfiltered after cooling to room temperature and allowing to stand forseveral days. It contains in 1 cc. 0.24006 g. of potassium acetatecorresponding to 21.1 per cent by weight.

5 cc. weigh 5.7033 g. (the density is 1.1407); after dilution with waterto 200 cc., 20 cc. require, in order to be neutralized againstphenolphthaleine, 37.50 cc. of n/5 KOH.

The halogen-free reaction product can be extracted from the reactionmass after the addition of water by means of ether; the etherealsolution is washed with bicarbonate and water and is dried with sodiumsulfate. After the evaporation of the solvent, there remains a crudecrystalline product which shows an unsharp melting point of 125-160 C.It represents a mixture of compounds which can be readily separated, byfractional sublimation in a high vacuum into an unsaturated ketone (35to 80%) and androsterone acetate (20 to 65%).

The androsterone acetate, after sublimating 011 the ketone, is depositedfrom 90 C. on at a pressure of mm. in the neck of the retort in welldeveloped druses of crystals and can be obtained from dilute alcohol inthe form of long needles of the melting point 160-161" C. It correspondsin all its properties with the androsterone acetate prepared fromnatural androsterone, does not show any melting point depression onmixing it with the latter and ex hibits the same optical rotation andthe same physiological activity. Yield The acetate was heated for 2hours with an excess of 3 n-methylalcoholic potassium hydroxidesolution; the reaction solution was diluted and extracted with ether.AIten-. evaporating the ether, the residue was carefully distilled at110 C. (10- mm.) and was subsequently recrystallized from dilutealcohol. Melting point 178 0. (not corrected). There were not found anydifferences on comparison with natural androsterone.

Example 2 As described in Example 1, first the unsaturated chloroketoneis produced. mg. of the same are placed with 100 mg. of potassiumbenzoate and 300 mg. of benzoic acid as solvent into a micro bomb tubeof 4 cm. length which then is closed by fusing, the mixture is heated ina glycerol bath for 20 minutes to about 200 C. and thereupon 100 minutesto about 150 C. (or altogether 90 minutes to 180 C.). An entirely clear,brown-red solution is obtained which solidifles on cooling. The contentof the bomb tube is dissolved in boiling alcohol, water is added, andthe aqueous solution is extracted with ether. The chlorine compoundremains in the aqueous solution while from the ethereal solution thebenzoate of the dehydro androsterone is isolated which is almostinsoluble in cold alcohol and which crystallizes from alcohol in prismsof the constant melting point 250 C. (not corrected).

The yield of the benzoic acid ester amounts on the aVerage to only 65%;the remaining portions are present in the form of unsaturated compounds.

The reaction of the chloroketone with potassium benzoate makes itpossible to precipitate the chloroketone of melting point 157 C.directly from the semicarbazone-crude decomposition products or from themother liquors in the form of the dehydro androsterone benzoate.

The benzoate of the dehydro androsterone of the melting point 250 C. canbe saponified by heating for 2 hours with 2 N-methylalcoholic potassiumhydroxide solution. On pouring the reaction mass into water and coolingwith ice, the dehydro androsterone precipitates directly in crystallineform; from dilute acetone ity is obtained in long 'white needles of theconstant melting point 148' C. (not corrected).

Dehydro androsterone proved to be effective in the caponcombtest; itsunit is about 600 7. It can be converted by hydrogenation into asaturated hydroxy ketone of 'the formula CmHauOz having a melting pointof 1'72 C., such compound being isomeric with the androsterone ofExample 1.

Of course, the given examples serve merely to illustrate the invention;various other modifications and changes in the processes and reagentsmay be made by those skilled in the art in accordance with theprinciples set forth herein and in the claims annexed hereto.

- What we claim is:

l. A method for the conversion of ketonic byproducts obtained onproducing the male sex hormone of the formula CmHaoOz, into the male sexhormone, said by-productshaving the carbon skeleton of the male sexhomone but having a double bond in the molecule and corre-. sponding tothe general formula CwHzaOz, said method comprising subjecting saidby-products to the action of a halogenizing agent, hydrogenating thedouble bond of the halogen compound obtained thereby under conditionswhich will not effect reduction of the keto group, reacting thehydrogenated halogen compound with an agent capable of replacing thehalogen with an acyloxy group, saponifying the ester compound to themale sex hormone so obtained, and isolating the latter.

2. A method for the conversion of ketonic byproducts obtained onproducing the male sex hormone of the formula CmHaoOa, into the esterderivatives of the male sex hormone, said byproducts having the carbonskeleton of the male sex hormone but having a double bond in themolecule and corresponding to the general formula C19H2a02, said methodcomprising subjecting said by-products to the action of a halogenizingagent, hydrogenating the double bond of the halogen compound obtainedthereby under conditions which will not efiect reduction of the ketogroup, reacting the hydrogenated halogen compound with an agent capableof replacing the halogen with an acyloxy group, and isolating the soobtained ester of the male sex hormone.

3. A method of producing the acid derivatives of the male sex hormone ofthe general formula 019113002, which comprises subjecting an unsaturatedcompound of the general formula CmHz-zOzX, wherein X represents an acidgroup,

-to the action of activated hydrogen to effect saturation of the nucleardouble bond, interrupting the hydrogenation procedure when 2 hydrogenatoms are absorbed, and isolating the saturated compound of the generalformula 019112902X.

4. A method for the conversion of ketonic byproducts obtained onproducing the male sex hormone into the male sex hormone and unsaturatedketo cyclopentano dimethyl polyhydrophenanthroles, said lay-productshaving the carbon skeleton of the male sex hormone but having a doublebond in the molecule and corresponding to the general formula CmHzaOz,said method said by-products into the male sex hormone and unsaturatedketo cyclopentano dimethyl polyhydro-phenanthroles, and separating. themale sex hormone from the latter.

5. A method for the production of the male sex hormone andketowyclopentanO-dimethylpolyhydro-phenanthroles from the ketonicbyproducts obtained on isolating the sex hormone from mixturescontaining the same, said byproducts having the carbon skeleton of themale sex hormone but having a double bond in the molecule andcorresponding to the general formula 0191 12802, said method beingcharacterized by the feature that the by-products are first transformedinto halogen compounds, which latter are converted by hydrogenation ofthe double bond in the molecule, reaction with an agent capable ofreplacing thehalogen with an acyloxy group, and saponification of theesters into the' male sex hormone, the hydrogenating, esterifying andsaponifying reactions being conducted in any order except that theesterification precedes the saponification, the hydrogenation beingconducted under such conditions that the keto group is not reduced,whereupon the hormone is separated from the unsaturatedketo-cyclopentano-dimethy1 polyhydro-phenanthroles which are alsoformed.

6. In a method for the conversion of ketonic by-products obtained onproducing the male sex hormone into a compound of the group consistingof the male sex hormone and other ketocyclopentano polyhydrophenanthrene compounds having a physiological action similar to that ofthe male sex hormone, said by-products having the carbon skeleton of themale sex hormone but having a double bond in the molecule andcorresponding to the general formula 0191-12802, the steps whichcomprise subjecting said by-products to hydrogenation and esterificationagents in any order, the hydrogenation agent being such as will saturatethe double bond in the molecule without reducing the keto group,

and the esterification agent being capable of introducing an acyloxygroup into the 3-position.

7. In a method for the conversion of ketonic by-products obtained onproducing the male sex hormone into a compound of the group consistingof the male sex hormone and other ketocyclopentano polyhydrophenanthrene compounds having a physiological action similar to that ofthe male sex hormone, said by-products having the carbon skeleton of themale sex hormone but having a double bond in the molecule andcorresponding to the general formula CmHzaOz, the steps which comprisesubjecting said by-products to halogenation, hydrogenation andesterification agents in any order except that esterification followshalogenation, the hydrogenation agent being such as will saturate thedouble bond in the molecule without reducing the keto group, and theesterification agent being capable of replacing halogen with an acyloxygroup.

8. In a method for the isolation of a male sex hormone and anunsaturated halogen ketone having the molecular structure of the malesex hormone from crude mixtures containing the male sex hormone and anunsaturated ketocompound having the carbon skeleton of the male sexhormone, the steps which comprise subjecting the crude starting materialcontaining the hormone to the action of a hydrolyzing and halogenatingagent of the group consisting of hydrogen halide and agents capable ofyielding hydrogen halide, and then separating the male sex hormone fromthe halogenated compound by distributing them between solvent mixturesin which they have different solubilities.

9. In a method of isolating the male sex hormone and an unsaturatedhalogen ketone having the molecular structure of the .male sex hormonefrom crude mixtures containing the male sex hormone and an unsaturatedketocompound having the carbon skeleton of the male sex hormone, thesteps of first subjecting the crude starting material containing thehormone to the action of a hydrolyzing and halogenating agent selectedfrom the group consisting of hydrogen halide and a compound capable ofyielding hydrogen halide, extracting said halogen ketone and male sexhormone by means of a hydrocarbon solvent, removing from said extractthe male sex hormone by treating the same with an aqueous,water-miscible solvent, the water content of which is so high that onlythe male sex hormone is removed, while the halogen ketone remains insolution, dissolving the halogen ketone from the hydrocarbon extract byextracting the same with an aqueous water-miscible solvent, the watercontent of which is so low that said unsaturated halogen ketone passesinto the water-miscible solvent, and isolating from the latter thehalogen ketone.

10. A method for the conversion of ketonic by-products obtained onproducing the male sex hormone, into a compound of the group consistingof the male sex hormone and other ketocyclopentano polyhydro penanthrenecompounds having a physiological action similar to that of the male sexhormone, said by-products having the structure of the male sex hormonebut having a double bond in the molecule and corresponding to thegeneral formula C19H2a02, said method comprising the steps of subjectingsaid by-products to hydrogenation under conditions which will effectsaturation of the double bond in the molecule but will not cause reduction of the keto group, esterification with an agent capable ofintroducing an acyloxy group into the 3-position, and saponification inany order except that the esterification precedes the saponification.

11. A cyclopentano phenanthrene derivative of the general formulaC19H29ZOR, wherein R is an acyl group and Z is a group obtained byreacting ketonic oxygen with a keto reagent.

12. A method for the production of the esters of the male sex hormonefrom ketonic compounds accompanying the hormone in mixtures obtained inthe production of the hormone, said compounds having the carbon skeletonof the male sex hormone but having a double bond in the molecule andcorresponding to the general formula C19H2a02, said method comprisingsubjecting said compounds to hydrogenation and esterification in anyorder, the hydrogenation agent being such as will saturate the doublebond in the molecule without reducing the keto group, and theesterification agent beingcapable of introducing an acyloxy group intothe 3- position, and isolating the saturated ester material.

13. A method for the production of the esters of the male sex hormonefrom halogenated ketonic compounds accompanying the hormone in mixturesobtained in the production of the hormone, said compounds having thegeneral formula C19H27OC1 and the carbon skeleton of the male sexhormone, which comprises subjecting said halogenated compounds tohydrogenation and esterification in any order, the hydrogenation agentbeing such as will saturate the double bond in the molecule withoutreducing the keto group, and the esterification agent being capable ofreplacing halogen with an acyloxy group, and isolating the saturatedester material.

14. The method according to claim 2, includ ing the step of reacting theketoacyloxy compound with a reagent capable of condensing with the ketogroup.

15. A process for saturating the nuclear double bond of a nuclearlyunsaturated keto cyclopentano-10,13-dimethvl polyhydro phenanthrenecompound, which comprises the hydrogenation of such double bond withhydrogen gas in the presence of a catalyst of the platinum metal group.

16. A process for saturating the nuclear double bond of a nuclearlyunsaturated keto cyclopentano-10,13-dimethyl polyhydro phenanthrenecompound, which comprises the hydrogenation of such double bond withhydrogen. gas in the presence of palladium.

17. A process for saturating the nuclear double bond of a nuclearlyunsaturated 3-substituted 17-keto cyclopentano-10,13-dimethyl polyhydrophenanthrene compound, which comprises the hydrogenation of such doublebond with hydrogen gas in the presence of a catalyst of the platinummetal group.

18. In a process for converting dehydro-androsterone into a saturatedderivative having the same carbon nucleus, the step which compriseseffecting saturation of the nuclear double bond with the aid of hydrogenin the presence of a catalyst of the platinum metal group.

19. In a process for converting dehydro-androsterone into a saturatedderivative having the same carbon nucleus, the step which comprisesefiecting saturation oi. the nuclear double bond with the aid ofhydrogen in the presence of palladium.

20. A process for the production of 3-hydroxyetioallo-cholanone-(l'l)from dehydro-androsterone which comprises the hydrogenation ofdehydroandrosterone by hydrogen gas in the resence of palladium.

21. The ketones of the formula CH CHI wherein R stands for an esterifiedhydroxyl group.-

22. The benzoate of dehydroandrosterone.

ADOLF BU'I'ENANDT.

